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Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Research Article

Peptide Specificity Analysis of Peptide: N-glycanases Using Synthetic Chitobiose-pentapeptides

Author(s): Taiki Kuribara , Toshihiro Ishihara , Takaya Kudo , Makoto Hirano and Kiichiro Totani*

Volume 24, Issue 8, 2017

Page: [723 - 728] Pages: 6

DOI: 10.2174/0929866524666170818160159

Price: $65

Abstract

Background: Peptide: N-glycanase is a deglycosylation enzyme releasing N-glycan from glycoproteins. Although glycan specificity analysis of this enzyme has been reported, recognition requirements for the peptide sequence have not been precisely elucidated.

Objective: In this study, we carried out peptide specificity analysis of several peptide:N-glycanases.

Methods: Using synthetic chitobiose-pentapeptide substrates having a systematic series of amino acid sequences composed of hydrophobic leucine and hydrophilic serine, we examined the peptide specificities of peptide: N-glycanases comprising yeast cytoplasmic PNGase, bacterial PNGase F, and plant PNGase A by ultra-performance liquid chromatography combined with electrospray ionization mass spectrometry.

Results: We found that each of the PNGases had higher activity for the more hydrophobic (leucinerich) chitobiose-pentapeptides, although the sensitivities of the PNGases for hydrophobicity varied. Cytoplasmic PNGase showed broad specificity. In contrast, PNGase A showed moderate specificity. PNGase F showed the highest specificity.

Conclusion: PNGases from different origins had similar but significantly independent peptide specificities.

Keywords: Peptide: N-glycanase, synthetic substrate, chitobiose-pentapeptide, peptide sequence, substrate specificity, ultraperformance liquid chromatography mass spectrometry.

Graphical Abstract

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